Department of Thoracic Surgery, Fudan University Cancer Hospital/Cancer Institute, Shanghai, PR China.
J Exp Clin Cancer Res. 2009 Aug 8;28(1):110. doi: 10.1186/1756-9966-28-110.
Despite multidisciplinary treatment, lung cancer remains a highly lethal disease due to poor response to chemotherapy. The identification of therapeutic agents with synergistic effects with traditional drugs is an alternative for lung cancer therapy. In this study, the synergistic effects of arsenic trioxide (As2O3) with cisplatin (DDP) on A549 and H460 non-small cell lung cancer (NSCLC) cells were explored.
A549 and H460 human lung cancer cells were treated with As2O3 and/or DDP. Cell growth curves, cell proliferation, cell cycle, and apoptosis of human cancer cell lines were determined by the 3-(4,5)-dimethylthiahiazo (-z-y1)-3,5-di-phenytetrazoliumromide (MTT) method, clonogenic assay, and flow cytometry (FCM). Apoptosis was further assessed by TUNEL staining. Cell cycle and apoptosis related protein p21, cyclin D1, Bcl-2, bax, clusterin, and caspase-3 were detected by western blot.
MTT and clonogenic assay showed As2O3 within 10(-2) microM to 10 microM exerted inhibition on the proliferation of NSCLC cells, and 2.5 microM As2O3 exerted synergistic inhibition on proliferation with 3 microg/ml DDP. The combination indices (CI) for A549 and H460 were 0.5 and 0.6, respectively, as confirmed by the synergism of As2O3 with DDP. FCM showed As2O3 did not affect the cell cycle. The G0/G1 fraction ranged from 57% to 62% for controlled A549 cells and cells treated with As2O3 and/or DDP. The G0/G1 fraction ranged from 37% to 42% for controlled H460 cells and cells treated with As2O3 and/or DDP. FCM and TUNEL staining illustrated that the combination of As2O3 and DDP provoked synergistic effects on apoptosis induction based on the analysis of the apoptosis index. Western blotting revealed that the expression of cell cycle related protein p21 and cyclin D1 were not affected by the treatments, whereas apoptosis related protein bax, Bcl-2, and clusterin were significantly regulated by As2O3 and/or DDP treatments compared with controls. The expression of caspase-3 in cells treated with the combination of As2O3 and DDP did not differ from that in cells treated with a single agent.
As2O3 exerted synergistic effects with DDP on NSCLC cells, and the synergistic effects were partly due to the induction of caspase-independent apoptosis.
尽管采用了多学科治疗，由于对化疗反应不佳，肺癌仍然是一种高度致命的疾病。寻找与传统药物具有协同作用的治疗剂是肺癌治疗的一种替代方法。在这项研究中，探讨了三氧化二砷（As2O3）与顺铂（DDP）联合应用对 A549 和 H460 非小细胞肺癌（NSCLC）细胞的协同作用。
用 As2O3 和/或 DDP 处理 A549 和 H460 人肺癌细胞。通过 3-(4,5)-二甲基噻唑 (-z-y1)-3,5-二苯基四唑溴盐（MTT）法、集落形成试验和流式细胞术（FCM）测定人癌细胞系的细胞生长曲线、细胞增殖、细胞周期和细胞凋亡。通过 TUNEL 染色进一步评估凋亡。通过 Western blot 检测细胞周期和凋亡相关蛋白 p21、细胞周期蛋白 D1、Bcl-2、bax、聚集素和 caspase-3。
MTT 和集落形成试验表明，As2O3 在 10(-2) microM 至 10 microM 范围内对 NSCLC 细胞的增殖有抑制作用，而 2.5 microM As2O3 与 3 µg/ml DDP 联合应用具有协同抑制增殖作用。As2O3 与 DDP 的协同作用证实了 A549 和 H460 的组合指数（CI）分别为 0.5 和 0.6。FCM 显示 As2O3 不影响细胞周期。对照 A549 细胞和用 As2O3 和/或 DDP 处理的细胞的 G0/G1 期范围为 57%至 62%。对照 H460 细胞和用 As2O3 和/或 DDP 处理的细胞的 G0/G1 期范围为 37%至 42%。FCM 和 TUNEL 染色表明，基于凋亡指数的分析，As2O3 和 DDP 的联合应用诱导了协同的细胞凋亡作用。Western blot 显示，细胞周期相关蛋白 p21 和细胞周期蛋白 D1 的表达不受处理的影响，而凋亡相关蛋白 bax、Bcl-2 和聚集素的表达在 As2O3 和/或 DDP 处理后与对照组相比明显受到调节。用 As2O3 和 DDP 联合处理的细胞中 caspase-3 的表达与用单一药物处理的细胞没有差异。
As2O3 与 DDP 对 NSCLC 细胞具有协同作用，协同作用部分归因于 caspase 非依赖性凋亡的诱导。
